- Email: [email protected]
Allicin inhibits spontaneous and TNF-fN˜induced secretion of proinflammatory cytokines and chemokines from intestinal epithelial cells
s44
EUROPEAN
Results: Mucosal atrophy (dilated intercellular spaces, cytoplasmic vesicu&ion, short microvilli, apoptotic nuclei, cell&u disorganisation) induced by PEM was less severe in pigs that consumed NAQ than in the rest of malnourished pigs. There were no differences in the ileal PPL subpopulations among malnourished groups. However, NAQ-fed pigs showed a closer PPL distribution to healthy pigs than the rest of malnourished pigs.
SOCIETY
OF PARENTERAL
AND
ENTERAL
NUTRlTION
Conclusions: Results presented in this study show that NAQ has a moderate protective effect, even better than Q, on the small intestinal damage induced by PEM
Method: Plasma IL-6 and CRP were measured pre- and for 5d post-op in 109 patients (61 males) undergoing surgery for gastrointestinal cancer. Genotyping by PCR and TaqMan techniques was performed. Results: The duration of operation and blood loss were gleater in males than females (duration 214f125 vs 172f76 min p=O.O32 Mann Whitney; blood loss 473f521 vs 258f348 ml p=O.O05 Mann Whitney) resulting in greater inflammatory stress. IL-6 and CRP concentrations peaked at 1 and 2 days post operatively, respectively in both sexes. Concentrations of IL-6 and CRP were greater in men, than women, on days 2 and 3, and day 4 respectively. Multivzuiate analysis, allowing for the duration of surgery and blood loss, showed IL-6 and CRP concentrations were greater in men with a TNF2 allele than women with the same genotype on day 2 postoperatively (554f494 vs lllf2lpg/ml and 193f116 vs 121f37mg/dl (piO.05 for both)). Other genotypes exerted no statistically significant influence. Conclusions: The inflammatory response in men but not women is upregulated by possession of polymorphisms in pro-inflammatory cytokine genes. This chxacteristic may impact upon post-operative morbidity and mortality.
163-P. IS HEALTHY AGEING ASSOCIATED OXIDANT-ANTIOXIDANT IMBALANCE ?
165-P. GLUTATHIONE METABOLISM IN HUMAN LIVER AFTER ISCHEMIA AND REPERFUSION
Table
1 Healthy
(x million
M-Control
M-Q
M-NAQ
cells)
CD4c
5.x *
CDS+
10.1 *
1.5
CD2lc
6.1 zk 1.4
1.7
2.9 zk 1.2 *
2.6 zk 0.8 *
4.5 * 0.8
5.3 *
5.0 *
1.7 *
6.1 zk 0.9
1.2 * 0.4 *
2.9 zk 0.6
1.0 *
1.7 * 0.7 *
W ITH AN
B.M. Winklhofer-Roob’, A. Meinitzer’, G. Khoschsoru?, M. Mxitschnegg’, W . Wonisch’, .I. Roob3, P. Grolier4, N. Cardinault4, B. Tiran’, I. Sundl’, G. Halwachs’, .I. Ribalta’, E. Rock4 ‘Inst M&c Biol, Biochem andMicrobio1, ‘Dept Lab Med, 3Div Clin Nephrol, Dept Internal Medicine, Karl-Franzens University, Graz, Austria, 41NRA, U M M M HNRC, Clermont-Ferrand, France, ‘Lipid Res Unit, Univ Rovira i Virgili, Reus, Spain Rationale: Oxidative stress is thought to play a crucial role in ageing. A major focus of the EU-funded VmAGE project is directed on the effects of healthy ageing on biomxkers of the oxidant-antioxidant imbalance. Method: For this purpose, we recorded dietxy intake and determined the exogenous antioxidants vitamin E, C, and cxotenoids in plasma, LDL, and buccal mucosal cells (BMC), and a selection of indices of lipid, protein and DNA oxidation, including LDL oxidizability, plasma total peroxide (POX) and malondialdehyde (MDA), minxy S-hydroxy-deoxyguanosine (S-OHDG) excretion in 300 healthy male non-smoking volunteers aged 20-75 yr and enrolled according to strict inclusion/exclusion criteria. Results: Dietxy intakes met the DRI and did not change with age. Plasma and LDL alpha-tocopherol (but not alpha-tocopherol:cholesterol) and lutein/zeaxanthin increased with age (PiO.OOl), while plasma and LDL lycopene and 1ycopene:cholesterol decreased (PiO.001) and vitamin C did not change. There were small but statistically significant increases with age in MDA (r=0.15, P=O.Ol) and POX (r=0.14, P=O.O2), while urinary S-OHDG and LDL oxidizability did not change. MDA and POX correlated with each other (r=0.40, PiO.001). Conclusions: Optimal antioxidant status seems to help maintain an oxidantantioxidant balance in the healthy elderly. Do they stay healthy because of good antioxidant supply or is only pathological ageing associated with severe oxidative stress?
164-P. CYTOKINE GENOTYPE AND GENDER INFLUENCE THE INFLAMMATORY RESPONSE T O SURGERY R.F. Grimble’, A. Thorell’, .I. Nygren’, 0. Ljungqvist’, N. Bxber’, S. Grant’, J. Madden’ ‘Institute of Human Nutrition, University of Southampton, Southampton, United Kingdom, ‘Centre for Gastrointestinal Disease, Ersta Hospital, Stockholm, Sweden Rationale: Cytokines have widespread metabolic effects in patients. Single nucleotide polymorphisms (SNP) in cytokine genes (e.g. TNF-alpha-308 (TNF2), TNF-“eta+252 (B22), IL-6 -174 (GG), IL-l”eta(-511 (CT/TT)) xe linked to a raised inflammatory response and poor clinical outcome. Men may be more likely to react adversely to surgery. The interaction between cytokine genotype and gender is unknown. W e examined this by studying changes in plasma IL-6 and CRP concentrations in relation to SNP in surgical patients.
B. Westman’, A. Th mel, 0. Rooyackers’, F. Hammarqvist’, .I. Werneiman’ ‘Dept of surgery, ‘Dept of aenestesiology, The Karolinska Institutet, Stockholm, Sweden Rationale: The liver is thougt be severely damaged after 20 minutes of &hernia. Despite this, liver resections due to malignancy are being done with &hernia times of up to 30 minutes in our hospital without any clinical signs of severe liver damage. This study was performed to elucidate whether &hernia and repel-fusion in the liver causes changes in glutathione metabolism as a sign of oxidative stress. Method: 6 patients with liver malignancy, scheduled for liver resection but otherwise healthy, were involved in the study. The mean age was 57 yezus, with an intelyal between 49 and 72 years. Liver biopsies were performed before &hernia, at maximal &hernia and at 5, 10, 15, 20, 25, and 30 minutes of repel-fusion. The biopsies were analysed for reduced (GSH) and total glutathione (tGSH). Oxidized glutathione (GSSG) was calculated as the difference between tGSH and GSH. Results: Values xe given as meanfSD. The &hernia times vzuied between 15 and 36 minutes with a mean of 28 minutes. Initial GSH level was 2.69f0.92.GSH then fluctuated during repelfusion between 3.18f1.08 and 2.79fO.SO.tGSH and GSSG vzuied in the same way and there was no change in ledox status of glutathione. The fluctuations were not significant. A peak was noted early postoperatively in transaminases of the liver with a mean of ASAT S.lf7.8 and ALAT S.Of7.9 respectively. Bilirubin was elevated to 68.5f42.0 and ALP to 21.3f40.0 Conclusions: Ischemia of about 30 minutes followed by reperfusion, does not exert oxidative stress in liver tissue, as measured by glutathione metabolism. These results implicates that compressing the portal vein and the hepatic artery could be extended to 30 minutes without risk of oxidative stress or irreversible liver damage.
166-P. ALLICIN INHIBITS SPONTANEOUS AND TNF-f INDUCED SECRETION O F PROINFLAMMATORY CYTOKINES AND CHEMOKINES F R O M INTESTINAL EPITHELIAL CELLS A. Lang, M. Lahav, S. Bx-Meir, Y. Chewers Gastroenterology, Sheba Medical Center, Tel Hashomer, Israel Rationale: Allicin, the active substance of fresh crushed garlic has different biological activities and was implicated as an anti-inflammatory agent. Epithelial cells have an important role in intestinal inflammation. The aim of this study was to assessthe immunomodulatory effect of allicin on intestinal epithelial cells Method: The spontaneous and TNF-alpha stimulated secretion of IL-lb&a, IL-S, IP-10 and MIG from HT-29 and Caco-2 cells was tested with or without pretreatment with allicin. Cytokine secretion was assessedusing ELISA and expression of mRNA was determined by an RNA protection assay
s45
CELL BIOLOGY, NUTRIENTS AND GENES
Results: Allicin mxkedly inhibited the spontaneous and TNF-alpha induced secretion of IL-lbeta, IL-S, IP-10 and MIG from the two different cell lines in a dose-dependent manner. Fmthelmore, allicin suppressed the expression of IL-S mRNA levels. No effect on the viability of the cells was noted. Allicin Inhibits Cytokine Secretion (HT.29)
No Allicin IL- lbeta IL-X
IP- 10 MIG
0.1*0.02 22&3.2
100s 30*4
Allicin 20 ( muM) 0.02*0.005 10*2 20&3 l&O.2
Conclusions: These observations indicate that allicin exerts an inhibitory immunomodulatory effect on intestinal epithelial cells and suggest that allicin may be used to attenuate intestinal inflammation.
167-P. EFFECTS OF ENDOTOXEMIA ON DIAPHRAGM MITOCHONDRIA IN 5 DAY VENTILATED PIGLETS K.S. Fred&son’, K. Ahlberg’, P. Radell’, L.I. E&son’, .I. Welnelman t , 0. Rooyackers t ‘Anaesthesiology and Intensive Care, Huddinge University Hospital, ‘Anaesthesiology and Intensive Care, Karolinska hospital, Karolinska Institutet, Stockholm, Sweden
169-P. SKELETAL
Rationale: Respiratory muscle weakness is a common problem in ICU patients. Previously we have shown that prolonged mechanical ventilation decreases mitochon&ial function in diaphragm muscle in piglets ventilated for 5 days (Clin Nutr 21, suppl 1, 69,2002). Here we investigate whether an additional endotoxaemia influences mitochondrial function and oxidative shess in diaphragm muscle. Method: Thirteen 2-4 month old piglets received controlled mechanical ventilation during 5 days. Six piglets received endotoxin during 3 horns on day 1. The endotoxin dose (5-10 wg/kg) was titrated to maintain pulmonary artery plessule equal to or larger then 60% of the systemic pressure. Diaphragm muscle biopsies wele obtained on day 5 and analysed for mitochon&ial enzymes (cytochrome c oxidase -COX- and citrate synthase -CS-) and superoxide dismutase (SOD) as an indicator of oxidative stress. Results: All control animals remained hemo-dynamically stable during the 5 days. One of the piglets in the endotoxin group died on the morning of day 5, while the other piglets where stabilised within two days. Table 1
Control (n=7) Endotoxin (n=6)
COX (~mol/min/g)
CS (~mol/min/g)
SOD (U/g)
44.1 * 9.7 39.6 zk 12.6
26.3 zk 5.X 25.6 zk 12.6
1923 zk 550 4109 * 1955s
Mean & SD. *: Significantly different from control (Student’s T-test).
Conclusions: Endotoxaemia had no effect on mavkers of diaphragm mitochondrial function in 5 days ventilated piglets despite an increased oxidative shess. These results indicate that also in ICU patients, prolonged respiratory support might influence diaphragm mitochon&ia more then the underlying disease.
168-P. MYOSTATIN MRNA EXPRESSION IS NOT ALTERED STREPTOZOTOCIN-INDUCED DIABETES AND INSULIN TREATMENT IN RAT SKELETAL MUSCLES
expression is not known. It is also not known whether myostatin expression is altered in catabolic insulin-deprived diabetic conditions Method: 12-week-old rats (8 per group) received one of the following treatmerits for eight weeks: 1) Streptozotocin-induced diabetes without treatment (DM); 2) Streptozotocin-induced diabetes with daily insulin treatment (Ultratard 10 U s.c.) (INS); 3) Non-diabetic control (C). Myostatin mRNA was measured (Northern Blot) in moderately oxidative gastrocnemius and highly oxidative soleus muscle Results: Plasma insulin was low in DM and nomalized by INS (C: 7.2f2.2; DM: O.Sf0.2; INS: 9.2f2.5 rig/ml). Body weight was also lower in DM and nex-normalized by INS (C: 422flO; DM: 306flO; INS: 394f6 grams; PiO.02: DM vs INS and C; PiO.05: INS vs C). Compxed to C, myostatin mRNA was unchanged in both DM and INS in gastrocnemius muscle (P>O.6). Myostatin mRNA was not detectable in C as well as DM and INS in soleus muscle Conclusions: Muscle-specific myostatin expression is not altered in insulindeprived diabetes under catabolic conditions. Insulin treatment also does not affect myostatin gene expression. A role of myostatin in loss of muscle mass in insulin-deprived diabetes is not supported by the current data. The cul7-ent study also does not support a role of basal insulin levels in the in viva regulation of myostatin gene expression
BY
R. Barazzoni ‘, A. Bosutti’ , M. Zanetti’ , G. Biolo’, M. Stebel’, L. Cattin ‘, G. Guamieri ’ ‘Dept of Clinical Morphological Technological Sciences Clinica Medica, ‘CSPA-Animal Facility, University of Trieste, Trieste, Italy Rationale: Myostatin negatively regulates muscle mass in animals and humans. Insulin is a principal anabolic hormone preserving muscle mass. Muscle mass is reduced during insulin deprivation in insulin-deprived diab&es. Whether insulin is involved in the in viva regulation of myostatin gene
MUSCLE APOPTOSIS IN GASTRIC CANCER
M. Bossola t , G. Doglietto ‘, F. Pacelli ‘, A. Broccolini’ , M. Muscxitoli3, D. Facchini3, A. Fava3, F. Rossi Fanelli3 ‘Surgery, ‘Neurology, Catholic University of Rome, 3Clinical Medicine, University ‘La Sapienza’, Rome, Italy Rationale: Muscle wasting in cancer is secondary to increased protein catabolism and/or reduced protein synthesis. Recently, it has been proposed that skeletal muscle apoptosis could occur in cancer and lead to muscle atrophy. Indeed skeletal muscle apoptosis is increased in malnourished patients affected by obstructive pulmonary disease or in patients with neuromuscular degenerative diseases. Data in cancer patients are lacking. The present study aimed at verifying whether skeletal muscle apoptosis is increased in gastric cancer patients Method: Six patients affected by gastric cancer and 2 patients affected by benign abdominal diseses candidate to laparotomy were included in the study. Rectus abdominis biopsies were obtained intraoperatively. Skeletal muscle apotosis was determined by fluorescent transferase-mediated dUTP nick-end labeling (TUNEL) assay and by immunohistochemishy for caspase 3. Results: Control patients were well-nourished (no weight loss and normal serum albumin levels). In patienst with gastric cancer, the stage of disease was II in one case, III in three patients and IV in two; mean preoperative serum albumin (gr/dL) levels wele 3.8+-0.4 and the mean weight loss (%) was 6.2+-1.6. In control patients and in 5 out of 6 of gastric cancer patients TUNEL assay and immunohistochemistry for caspase 3 were negative. In one patien with gastric cancer TUNEL positive nuclei were observed (5 1% of all examined nuclei). Conclusions: These preliminary results would suggest that skeletal muscle apoptosis does not occur in moderately malnourished gashic cancer patients.
170-P. INCREASED SOLUBLE INTERLEUKIN-2 CONCENTRATIONS IN PATIENTS RECEIVING POSTOPERATIVE ENTERAL NUTRITION
RECEPTOR EARLY
M.E.P. Wir n ‘, .I. Emerudh’, .I. Pelmert ‘, .I. Larson ’ ’ Center for Surgical Sciences, Karolinska Institute, Stockholm, ‘Department of Immunology, Linkoping University, Linkoping, Sweden Rationale: Early enteral feeding improves gut integrity postoperatively and can reduce infectious complications. The aim of this study was to compare the effects of enteral feeding to parentera nutrition after major abdominal surgery on immunology, protein metabolism and clinical outcome. Method: Patients scheduled for major GI surgery were randomly allocated to enteral (EN) or pxenteral nutrition (PN) for five days, n=20. The nu-
EUROPEAN
Results: Mucosal atrophy (dilated intercellular spaces, cytoplasmic vesicu&ion, short microvilli, apoptotic nuclei, cell&u disorganisation) induced by PEM was less severe in pigs that consumed NAQ than in the rest of malnourished pigs. There were no differences in the ileal PPL subpopulations among malnourished groups. However, NAQ-fed pigs showed a closer PPL distribution to healthy pigs than the rest of malnourished pigs.
SOCIETY
OF PARENTERAL
AND
ENTERAL
NUTRlTION
Conclusions: Results presented in this study show that NAQ has a moderate protective effect, even better than Q, on the small intestinal damage induced by PEM
Method: Plasma IL-6 and CRP were measured pre- and for 5d post-op in 109 patients (61 males) undergoing surgery for gastrointestinal cancer. Genotyping by PCR and TaqMan techniques was performed. Results: The duration of operation and blood loss were gleater in males than females (duration 214f125 vs 172f76 min p=O.O32 Mann Whitney; blood loss 473f521 vs 258f348 ml p=O.O05 Mann Whitney) resulting in greater inflammatory stress. IL-6 and CRP concentrations peaked at 1 and 2 days post operatively, respectively in both sexes. Concentrations of IL-6 and CRP were greater in men, than women, on days 2 and 3, and day 4 respectively. Multivzuiate analysis, allowing for the duration of surgery and blood loss, showed IL-6 and CRP concentrations were greater in men with a TNF2 allele than women with the same genotype on day 2 postoperatively (554f494 vs lllf2lpg/ml and 193f116 vs 121f37mg/dl (piO.05 for both)). Other genotypes exerted no statistically significant influence. Conclusions: The inflammatory response in men but not women is upregulated by possession of polymorphisms in pro-inflammatory cytokine genes. This chxacteristic may impact upon post-operative morbidity and mortality.
163-P. IS HEALTHY AGEING ASSOCIATED OXIDANT-ANTIOXIDANT IMBALANCE ?
165-P. GLUTATHIONE METABOLISM IN HUMAN LIVER AFTER ISCHEMIA AND REPERFUSION
Table
1 Healthy
(x million
M-Control
M-Q
M-NAQ
cells)
CD4c
5.x *
CDS+
10.1 *
1.5
CD2lc
6.1 zk 1.4
1.7
2.9 zk 1.2 *
2.6 zk 0.8 *
4.5 * 0.8
5.3 *
5.0 *
1.7 *
6.1 zk 0.9
1.2 * 0.4 *
2.9 zk 0.6
1.0 *
1.7 * 0.7 *
W ITH AN
B.M. Winklhofer-Roob’, A. Meinitzer’, G. Khoschsoru?, M. Mxitschnegg’, W . Wonisch’, .I. Roob3, P. Grolier4, N. Cardinault4, B. Tiran’, I. Sundl’, G. Halwachs’, .I. Ribalta’, E. Rock4 ‘Inst M&c Biol, Biochem andMicrobio1, ‘Dept Lab Med, 3Div Clin Nephrol, Dept Internal Medicine, Karl-Franzens University, Graz, Austria, 41NRA, U M M M HNRC, Clermont-Ferrand, France, ‘Lipid Res Unit, Univ Rovira i Virgili, Reus, Spain Rationale: Oxidative stress is thought to play a crucial role in ageing. A major focus of the EU-funded VmAGE project is directed on the effects of healthy ageing on biomxkers of the oxidant-antioxidant imbalance. Method: For this purpose, we recorded dietxy intake and determined the exogenous antioxidants vitamin E, C, and cxotenoids in plasma, LDL, and buccal mucosal cells (BMC), and a selection of indices of lipid, protein and DNA oxidation, including LDL oxidizability, plasma total peroxide (POX) and malondialdehyde (MDA), minxy S-hydroxy-deoxyguanosine (S-OHDG) excretion in 300 healthy male non-smoking volunteers aged 20-75 yr and enrolled according to strict inclusion/exclusion criteria. Results: Dietxy intakes met the DRI and did not change with age. Plasma and LDL alpha-tocopherol (but not alpha-tocopherol:cholesterol) and lutein/zeaxanthin increased with age (PiO.OOl), while plasma and LDL lycopene and 1ycopene:cholesterol decreased (PiO.001) and vitamin C did not change. There were small but statistically significant increases with age in MDA (r=0.15, P=O.Ol) and POX (r=0.14, P=O.O2), while urinary S-OHDG and LDL oxidizability did not change. MDA and POX correlated with each other (r=0.40, PiO.001). Conclusions: Optimal antioxidant status seems to help maintain an oxidantantioxidant balance in the healthy elderly. Do they stay healthy because of good antioxidant supply or is only pathological ageing associated with severe oxidative stress?
164-P. CYTOKINE GENOTYPE AND GENDER INFLUENCE THE INFLAMMATORY RESPONSE T O SURGERY R.F. Grimble’, A. Thorell’, .I. Nygren’, 0. Ljungqvist’, N. Bxber’, S. Grant’, J. Madden’ ‘Institute of Human Nutrition, University of Southampton, Southampton, United Kingdom, ‘Centre for Gastrointestinal Disease, Ersta Hospital, Stockholm, Sweden Rationale: Cytokines have widespread metabolic effects in patients. Single nucleotide polymorphisms (SNP) in cytokine genes (e.g. TNF-alpha-308 (TNF2), TNF-“eta+252 (B22), IL-6 -174 (GG), IL-l”eta(-511 (CT/TT)) xe linked to a raised inflammatory response and poor clinical outcome. Men may be more likely to react adversely to surgery. The interaction between cytokine genotype and gender is unknown. W e examined this by studying changes in plasma IL-6 and CRP concentrations in relation to SNP in surgical patients.
B. Westman’, A. Th mel, 0. Rooyackers’, F. Hammarqvist’, .I. Werneiman’ ‘Dept of surgery, ‘Dept of aenestesiology, The Karolinska Institutet, Stockholm, Sweden Rationale: The liver is thougt be severely damaged after 20 minutes of &hernia. Despite this, liver resections due to malignancy are being done with &hernia times of up to 30 minutes in our hospital without any clinical signs of severe liver damage. This study was performed to elucidate whether &hernia and repel-fusion in the liver causes changes in glutathione metabolism as a sign of oxidative stress. Method: 6 patients with liver malignancy, scheduled for liver resection but otherwise healthy, were involved in the study. The mean age was 57 yezus, with an intelyal between 49 and 72 years. Liver biopsies were performed before &hernia, at maximal &hernia and at 5, 10, 15, 20, 25, and 30 minutes of repel-fusion. The biopsies were analysed for reduced (GSH) and total glutathione (tGSH). Oxidized glutathione (GSSG) was calculated as the difference between tGSH and GSH. Results: Values xe given as meanfSD. The &hernia times vzuied between 15 and 36 minutes with a mean of 28 minutes. Initial GSH level was 2.69f0.92.GSH then fluctuated during repelfusion between 3.18f1.08 and 2.79fO.SO.tGSH and GSSG vzuied in the same way and there was no change in ledox status of glutathione. The fluctuations were not significant. A peak was noted early postoperatively in transaminases of the liver with a mean of ASAT S.lf7.8 and ALAT S.Of7.9 respectively. Bilirubin was elevated to 68.5f42.0 and ALP to 21.3f40.0 Conclusions: Ischemia of about 30 minutes followed by reperfusion, does not exert oxidative stress in liver tissue, as measured by glutathione metabolism. These results implicates that compressing the portal vein and the hepatic artery could be extended to 30 minutes without risk of oxidative stress or irreversible liver damage.
166-P. ALLICIN INHIBITS SPONTANEOUS AND TNF-f INDUCED SECRETION O F PROINFLAMMATORY CYTOKINES AND CHEMOKINES F R O M INTESTINAL EPITHELIAL CELLS A. Lang, M. Lahav, S. Bx-Meir, Y. Chewers Gastroenterology, Sheba Medical Center, Tel Hashomer, Israel Rationale: Allicin, the active substance of fresh crushed garlic has different biological activities and was implicated as an anti-inflammatory agent. Epithelial cells have an important role in intestinal inflammation. The aim of this study was to assessthe immunomodulatory effect of allicin on intestinal epithelial cells Method: The spontaneous and TNF-alpha stimulated secretion of IL-lb&a, IL-S, IP-10 and MIG from HT-29 and Caco-2 cells was tested with or without pretreatment with allicin. Cytokine secretion was assessedusing ELISA and expression of mRNA was determined by an RNA protection assay
s45
CELL BIOLOGY, NUTRIENTS AND GENES
Results: Allicin mxkedly inhibited the spontaneous and TNF-alpha induced secretion of IL-lbeta, IL-S, IP-10 and MIG from the two different cell lines in a dose-dependent manner. Fmthelmore, allicin suppressed the expression of IL-S mRNA levels. No effect on the viability of the cells was noted. Allicin Inhibits Cytokine Secretion (HT.29)
No Allicin IL- lbeta IL-X
IP- 10 MIG
0.1*0.02 22&3.2
100s 30*4
Allicin 20 ( muM) 0.02*0.005 10*2 20&3 l&O.2
Conclusions: These observations indicate that allicin exerts an inhibitory immunomodulatory effect on intestinal epithelial cells and suggest that allicin may be used to attenuate intestinal inflammation.
167-P. EFFECTS OF ENDOTOXEMIA ON DIAPHRAGM MITOCHONDRIA IN 5 DAY VENTILATED PIGLETS K.S. Fred&son’, K. Ahlberg’, P. Radell’, L.I. E&son’, .I. Welnelman t , 0. Rooyackers t ‘Anaesthesiology and Intensive Care, Huddinge University Hospital, ‘Anaesthesiology and Intensive Care, Karolinska hospital, Karolinska Institutet, Stockholm, Sweden
169-P. SKELETAL
Rationale: Respiratory muscle weakness is a common problem in ICU patients. Previously we have shown that prolonged mechanical ventilation decreases mitochon&ial function in diaphragm muscle in piglets ventilated for 5 days (Clin Nutr 21, suppl 1, 69,2002). Here we investigate whether an additional endotoxaemia influences mitochondrial function and oxidative shess in diaphragm muscle. Method: Thirteen 2-4 month old piglets received controlled mechanical ventilation during 5 days. Six piglets received endotoxin during 3 horns on day 1. The endotoxin dose (5-10 wg/kg) was titrated to maintain pulmonary artery plessule equal to or larger then 60% of the systemic pressure. Diaphragm muscle biopsies wele obtained on day 5 and analysed for mitochon&ial enzymes (cytochrome c oxidase -COX- and citrate synthase -CS-) and superoxide dismutase (SOD) as an indicator of oxidative stress. Results: All control animals remained hemo-dynamically stable during the 5 days. One of the piglets in the endotoxin group died on the morning of day 5, while the other piglets where stabilised within two days. Table 1
Control (n=7) Endotoxin (n=6)
COX (~mol/min/g)
CS (~mol/min/g)
SOD (U/g)
44.1 * 9.7 39.6 zk 12.6
26.3 zk 5.X 25.6 zk 12.6
1923 zk 550 4109 * 1955s
Mean & SD. *: Significantly different from control (Student’s T-test).
Conclusions: Endotoxaemia had no effect on mavkers of diaphragm mitochondrial function in 5 days ventilated piglets despite an increased oxidative shess. These results indicate that also in ICU patients, prolonged respiratory support might influence diaphragm mitochon&ia more then the underlying disease.
168-P. MYOSTATIN MRNA EXPRESSION IS NOT ALTERED STREPTOZOTOCIN-INDUCED DIABETES AND INSULIN TREATMENT IN RAT SKELETAL MUSCLES
expression is not known. It is also not known whether myostatin expression is altered in catabolic insulin-deprived diabetic conditions Method: 12-week-old rats (8 per group) received one of the following treatmerits for eight weeks: 1) Streptozotocin-induced diabetes without treatment (DM); 2) Streptozotocin-induced diabetes with daily insulin treatment (Ultratard 10 U s.c.) (INS); 3) Non-diabetic control (C). Myostatin mRNA was measured (Northern Blot) in moderately oxidative gastrocnemius and highly oxidative soleus muscle Results: Plasma insulin was low in DM and nomalized by INS (C: 7.2f2.2; DM: O.Sf0.2; INS: 9.2f2.5 rig/ml). Body weight was also lower in DM and nex-normalized by INS (C: 422flO; DM: 306flO; INS: 394f6 grams; PiO.02: DM vs INS and C; PiO.05: INS vs C). Compxed to C, myostatin mRNA was unchanged in both DM and INS in gastrocnemius muscle (P>O.6). Myostatin mRNA was not detectable in C as well as DM and INS in soleus muscle Conclusions: Muscle-specific myostatin expression is not altered in insulindeprived diabetes under catabolic conditions. Insulin treatment also does not affect myostatin gene expression. A role of myostatin in loss of muscle mass in insulin-deprived diabetes is not supported by the current data. The cul7-ent study also does not support a role of basal insulin levels in the in viva regulation of myostatin gene expression
BY
R. Barazzoni ‘, A. Bosutti’ , M. Zanetti’ , G. Biolo’, M. Stebel’, L. Cattin ‘, G. Guamieri ’ ‘Dept of Clinical Morphological Technological Sciences Clinica Medica, ‘CSPA-Animal Facility, University of Trieste, Trieste, Italy Rationale: Myostatin negatively regulates muscle mass in animals and humans. Insulin is a principal anabolic hormone preserving muscle mass. Muscle mass is reduced during insulin deprivation in insulin-deprived diab&es. Whether insulin is involved in the in viva regulation of myostatin gene
MUSCLE APOPTOSIS IN GASTRIC CANCER
M. Bossola t , G. Doglietto ‘, F. Pacelli ‘, A. Broccolini’ , M. Muscxitoli3, D. Facchini3, A. Fava3, F. Rossi Fanelli3 ‘Surgery, ‘Neurology, Catholic University of Rome, 3Clinical Medicine, University ‘La Sapienza’, Rome, Italy Rationale: Muscle wasting in cancer is secondary to increased protein catabolism and/or reduced protein synthesis. Recently, it has been proposed that skeletal muscle apoptosis could occur in cancer and lead to muscle atrophy. Indeed skeletal muscle apoptosis is increased in malnourished patients affected by obstructive pulmonary disease or in patients with neuromuscular degenerative diseases. Data in cancer patients are lacking. The present study aimed at verifying whether skeletal muscle apoptosis is increased in gastric cancer patients Method: Six patients affected by gastric cancer and 2 patients affected by benign abdominal diseses candidate to laparotomy were included in the study. Rectus abdominis biopsies were obtained intraoperatively. Skeletal muscle apotosis was determined by fluorescent transferase-mediated dUTP nick-end labeling (TUNEL) assay and by immunohistochemishy for caspase 3. Results: Control patients were well-nourished (no weight loss and normal serum albumin levels). In patienst with gastric cancer, the stage of disease was II in one case, III in three patients and IV in two; mean preoperative serum albumin (gr/dL) levels wele 3.8+-0.4 and the mean weight loss (%) was 6.2+-1.6. In control patients and in 5 out of 6 of gastric cancer patients TUNEL assay and immunohistochemistry for caspase 3 were negative. In one patien with gastric cancer TUNEL positive nuclei were observed (5 1% of all examined nuclei). Conclusions: These preliminary results would suggest that skeletal muscle apoptosis does not occur in moderately malnourished gashic cancer patients.
170-P. INCREASED SOLUBLE INTERLEUKIN-2 CONCENTRATIONS IN PATIENTS RECEIVING POSTOPERATIVE ENTERAL NUTRITION
RECEPTOR EARLY
M.E.P. Wir n ‘, .I. Emerudh’, .I. Pelmert ‘, .I. Larson ’ ’ Center for Surgical Sciences, Karolinska Institute, Stockholm, ‘Department of Immunology, Linkoping University, Linkoping, Sweden Rationale: Early enteral feeding improves gut integrity postoperatively and can reduce infectious complications. The aim of this study was to compare the effects of enteral feeding to parentera nutrition after major abdominal surgery on immunology, protein metabolism and clinical outcome. Method: Patients scheduled for major GI surgery were randomly allocated to enteral (EN) or pxenteral nutrition (PN) for five days, n=20. The nu-